ASTM F2386-04
(Guide)Standard Guide for Preservation of Tissue Engineered Medical Products (TEMPs) (Withdrawn 2013)
Standard Guide for Preservation of Tissue Engineered Medical Products (TEMPs) (Withdrawn 2013)
SIGNIFICANCE AND USE
The preservation of TEMPs can affect their subsequent characteristics (for example, structural, mechanical, biological, and metabolic properties). The aspects of preservation that can most adversely affect these characteristics include, but are not limited to, cooling, cooling rates, warming, warming rates, freezing, freezing rates, thawing, thawing rates, preservation medium, and pre- and post-processing.
The intent of this guide is to outline procedures that can minimize the loss or degradation of the desired characteristics of the TEMPs.
SCOPE
1.1 This guide covers the development of standards related to the preservation of cells, tissues, and tissue engineered medical products (TEMPs). Preservation techniques include freezing, vitrification, and hypothermic preservation. This guide discusses preservation, including issues of pre-preservation processing, the process of preservation, storage, transport, recovery, post-preservation processing, quality assurance, and process control.
1.2 This guide contains general guidelines for the preservation of cells, tissues, and tissue engineered medical products (TEMPs) and will identify more specific parameters relevant to the preservation of TEMPs.
1.3 This guide does not apply to any medical products of human origin regulated by the U.S. Food and Drug Administration (FDA) under 21 CFR Parts 16 and 1270 and 21 CFR Parts 207, 807, and 1271.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory requirements prior to use.
WITHDRAWN RATIONALE
This guide covers the development of standards related to the preservation of cells, tissues, and tissue engineered medical products (TEMPs). Preservation techniques include freezing, vitrification, and hypothermic preservation. This guide discusses preservation, including issues of pre-preservation processing, the process of preservation, storage, transport, recovery, post-preservation processing, quality assurance, and process control.
Formerly under the jurisdiction of Committee F04 on Medical and Surgical Materials and Devices, this guide was withdrawn in January 2013 in accordance with section 10.5.3.1 of the Regulations Governing ASTM Technical Committees, which requires that standards shall be updated by the end of the eighth year since the last approval date.
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Standards Content (Sample)
NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: F2386 − 04
StandardGuide for
Preservation of Tissue Engineered Medical Products
(TEMPs)
This standard is issued under the fixed designation F2386; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
The ability to preserve an engineered tissue or cell-based therapeutic device is important for its
clinicalapplication.Specifically,preservationpermitstheabilitytostoreandtransporttheproduct,the
time for the completion of safety and quality control testing prior to use in patients, and the ability to
uncouplethetimingofproductionoftheengineeredproductfromitsuse,ensuringadequateinventory
control and the ability to avoid both shortages and product expiration. The aim of this guide is to
identify key parameters relevant to the preservation of engineered tissues and cell-based therapies.
1. Scope 2. Referenced Documents
2.1 ASTM Standards:
1.1 This guide covers the development of standards related
E1564 Guide for Design and Maintenance of Low-
to the preservation of cells, tissues, and tissue engineered
Temperature Storage Facilities for Maintaining Cryopre-
medical products (TEMPs). Preservation techniques include
served Biological Materials
freezing, vitrification, and hypothermic preservation. This
E1565Guide for Inventory Control and Handling of Bio-
guide discusses preservation, including issues of pre-
logical Material Maintained at Low Temperatures
preservation processing, the process of preservation, storage,
2.2 Code of Federal Regulations:
transport, recovery, post-preservation processing, quality
21 CFR 610.18Standards for Establishment of Master Cell
assurance, and process control.
Banks
1.2 This guide contains general guidelines for the preserva-
21 CFR Parts 16 and 1270Human Tissue Intended for
tion of cells, tissues, and tissue engineered medical products
Transplantation
(TEMPs)andwillidentifymorespecificparametersrelevantto
21CFRParts207,807,and1271HumanCells,Tissues,and
the preservation of TEMPs.
Cellular and Tissue-Based Products: Establishment Reg-
istration and Listing
1.3 This guide does not apply to any medical products of
2.3 American Association of Blood Banks, Standards:
human origin regulated by the U.S. Food and Drug Adminis-
Preparation of Blood Components
tration (FDA) under 21 CFR Parts 16 and 1270 and 21 CFR
Conditions for Storage, Transportation and Expiration
Parts 207, 807, and 1271.
Tissue Storage and Issue
1.4 This standard does not purport to address all of the
2.4 American Association of Blood Banks, AABB Technical
safety concerns, if any, associated with its use. It is the
Manual, Methods:
responsibility of the user of this standard to establish appro- Method 6.18Cryopreservation of Hematopoietic Progenitor
priate safety and health practices and determine the applica-
Cells Using a Controlled Rate Freeze
bility of regulatory requirements prior to use.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on
This guide is under the jurisdiction of ASTM Committee F04 on Medical and the ASTM website.
Surgical Materials and Devicesand is the direct responsibility of Subcommittee AvailablefromU.S.GovernmentPrintingOfficeSuperintendentofDocuments,
F04.45 on Adventitious Agents Safety. 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401.
Current edition approved June 1, 2004. Published June 2004. DOI: 10.1520/ Available from American Association of Blood Banks (AABB), 8101 Glen-
F2386-04. brook Rd., Bethesda, MD 20814-2749.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F2386 − 04
Method 7.2Testing Freezer Alarms (3) lyophilization—in common usage, a synonym for
Method 7.3Monitoring Temperature During Shipment freeze-drying. Whereas etymologically, this term also com-
prises other methods for anhydrobiotic preservation, care
3. Terminology should be taken to avoid ambiguity.
3.1.12.2 cryopreservation—preservation by cooling to a
3.1 Definitions:
temperature below the equilibrium freezing temperature of the
3.1.1 cooling rate—absolute value of the instantaneous rate
preservation solution, such that there is solidification. The
of change of temperature during cooling.
resulting solid may be either crystalline or amorphous, or a
3.1.2 cryopreservation solution—a preservation medium to
combination of crystalline and amorphous phases.
which has been added one or more cryoprotectants.
(1) freezing—cryopreservation by formation of crystals.
3.1.3 cryoprotectant—a chemical or biological substance or
Frozen cell suspensions or TEMPs typically contain both
mixture of substances used to protect cells or matrix, or both,
crystalline and amorphous water.
during cryopreservation and rewarming. In general usage, a
(2) vitrification—cryopreservation by formation of glass.
cryoprotectant is added to a preservation medium to form a
Vitrified cell suspensions or TEMPs may contain small
cryopreservation solution.
amounts of crystalline water.
3.1.4 cytocrit—the ratio of cell volume to the total volume
3.1.12.3 hypothermic preservation—preservation by cool-
of solution and cells for a cell suspension.
ing to any temperature below normothermic culture
temperatures, such that biological components are suspended
3.1.5 dry shipper—a storage and transportation device for
in a liquid phase. Hypothermic storage temperatures may be
frozen products that contains a liquid nitrogen (LN) absorbent
either above or below the equilibrium freezing temperature.
material in the walls of the container. This device is designed
to maintain cryogenic temperature for several days.
3.1.13 preserve—to stabilize for the purposes of maintain-
ingthespecificmechanical,structural,metabolic,orbiological
3.1.6 equilibrium freezing temperature—temperature at
characteristics.
which an aqueous solution of a given composition is in
equilibrium with ice.
3.1.14 post-preservation processing—manipulation of cells
3.1.7 eutectic point—a temperature(s) at which the solute(s) or tissue after completion of the preservation process. These
in a solution become saturated due to freeze-concentration and steps include, but are not limited to, washing of the cells or
precipitatefromthesolution,causingtheunfrozenliquidwater tissue to remove the preservation medium/solution and/or
to simultaneously freeze. specific chemical or biological agents; dilution or concentra-
tion of the cells or tissue; and warming to temperatures
3.1.8 glass transition temperature—the temperature at
appropriate for the normal or desired physiological or bio-
whichtheheatcapacityassociatedwithtranslationalmolecular
chemical functions, or both, of the cells or tissue.
motions vanishes during cooling or appears during warming.
The glass transition temperature is the formal transition point
3.1.15 reconstitute—to add a solvent or diluent to an anhy-
between the glassy state and the liquid state.
drobiotically preserved sample in order to dissolve or suspend
its components.
3.1.9 nucleation—the formation of ice from a supercooled
aqueous solution by random aggregation of water molecules
3.1.16 rewarm—towarmfrompreservationtemperaturetoa
into clusters with ice-like properties. Nucleation may be
temperaturerequiredforuse(forexample,additionalcultureor
homogenous (spontaneous) or heterogenous (catalyzed by a
clinical use).
substrate that reduces the thermodynamic barrier to cluster
3.1.17 seeding—the deliberate initiation of ice crystal for-
formation).
mation in a supercooled aqueous solution under controlled
3.1.10 pre-preservation processing—the manipulation of
conditions. When ice forms either spontaneously or as a result
cellsortissuepriortotheinitiationofpreservation.Thesesteps
of the inclusion of nucleating agents in the cryopreservation
include, but are not limited to, selection of specific cell
solution, this is referred to as nucleation.
populations for freezing, centrifugation to modify cell density,
3.1.18 storage temperature—temperatureatwhichthecells,
introduction of specific chemical or biological agents, and
cooling. tissue,orTEMPisheldaftercompletionofthecoolingprocess.
3.1.11 preservation medium—a specific formulation of an
3.1.19 supercool—to cool to a temperature below the equi-
aqueous or nonaqueous solution in which a population of cells
librium melting point of the solution without initiating ice
or a tissue will be preserved.
formation.
3.1.12 preservation technologies:
3.1.20 temperature profile—the time-temperature history of
3.1.12.1 anhydrobiotic preservation—preservation by va-
a sample during cooling or warming.
porization and removal of water.
3.1.21 thaw—to warm from a cryopreserved state to a
(1) desiccation—a process for anhydrobiotic preservation
temperature above the melting point of the preservation me-
in which water is removed by evaporation.
dium.
(2) freeze-drying—aprocessforanhydrobioticpreservation
in which ice crystals are formed by freezing, and water is 3.1.22 warming rate—instantaneous rate of change of tem-
removed by sublimation and evaporation. perature during warming.
F2386 − 04
4. Summary of Guide development of selection protocols to prevent additional
stresses to the cells such as nutrient or oxygen deprivation that
4.1 The preservation of TEMPs can affect their subsequent
maycompromisetheirabilitytosurvivethestressesoffreezing
characteristics(forexample,structural,mechanical,biological,
and thawing.
and metabolic properties).The aspects of preservation that can
most adversely affect these characteristics include, but are not 5.4 Hypothermic Preservation:
limited to, cooling, cooling rates, warming, warming rates, 5.4.1 Preservation—A protocol for hypothermic storage
freezing, freezing rates, thawing, thawing rates, preservation will typically specify a temperature (or range of temperatures),
medium, and pre- and post-processing. cell concentration (for cell suspensions), composition of pres-
ervationsolution,maximumdurationofstorage,andmethodof
4.2 The intent of this guide is to outline procedures that can
use (for example, under perfusion or static conditions). Pres-
minimize the loss or degradation of the desired characteristics
ervation solutions used for hypothermic storage are typically
of the TEMPs.
designed to meet the metabolic needs of the cells (oxygen and
nutrients), prevent cell swelling and stabilize the cytoskeleton
5. Procedure
during storage.
5.1 Inventory Control and Handling—The methods for
5.4.2 Storage—Storage units must be designed to maintain
product tracking and identification during preservation must
the appropriate temperature throughout the storage area.
conform to methods developed for the manufacturing process
5.4.2.1 Storage units shall have alarm systems with audible
of the product. Materials for labels and labeling must be
signals.
compatible with the preservation process. For example, labels
5.4.2.2 The alarm must be set to activate at a temperature
for cryopreserved products must remain adherent at the appro-
that will allow for proper action to be taken before the product
priate storage temperatures or must be sealed into a pouch
has reached a temperature that makes the product unusable.
connected to the product container. Adhesives or other mate-
5.4.2.3 The alarm must signal in an area with adequate
rials used for the label must not leach into the product.
personnel to ensure that immediate corrective action will be
taken.
5.2 Preliminary Processing—In contrast to native tissues,
TEMPs may require extensive periods of ex vivo culture. The 5.4.2.4 Written procedures must be available containing
directions on methods to maintain the cells, tissues, or TEMPs
cultureconditions(compositionofmedium,oxygen,andwaste
concentrations) can influence cell metabolism and membrane within permissible temperatures and instructions to be fol-
lowed in the event of power failure.
composition. These factors can influence the ability of cells to
be successfully cryopreserved. To date, vascularized TEMPs 5.4.3 Transportation—A range of acceptable temperatures
forshipmentmustbeestablished.Somemethodoftemperature
have not been developed. The lack of vasculature will influ-
ence the flow of water and propagation of ice during cryo- monitoring is desirable when samples are shipped.
5.4.4 Post-transit Inspection—The sample and shipping
preservation.Thus,preservationprotocolsdevelopedfornative
tissues may not be appropriate for TEMPs designed to replace container should be inspected upon receipt. The sample and
thesametissue(forexample,skin).Coolingofcellsandtissues sample identifier should correspond to the intended product.
can lead to loss in cellular viability during the cooling process The sample and its container should be free of cracks or other
and/or sensitize cells or tissues to preservation at cryogenic externalevidenceofdamage.Protocolsfordecontaminationof
temperaturessuchthatviabilityorfunction,orboth,arealtered the shipping container in case of sample breakage should also
following a return to a desired physiological state. For be available.
example, during cooling, specific enzymatic functions associ-
5.5 Freezing:
ated with ion transport can become impaired, resulting in an
5.5.1 Prepreservation Processings: Osmotic
inability of the cells to maintain ion balance, leading to
Considerations—Additives used to protect cells upon freezing
subsequent loss in cellular viability or function, or both. Effort
and rewarming alter the osmolality of the solution. The
should be made to determine the sensitivity of the cells or
introductionofcellsortissuestypicallyculturedunderisotonic
tissues to be preserved to the effects of cooling prior to
conditions into a hypertonic preservation solution can result in
preservation at cryogenic temperatures.
a rapid efflux of water from the cell followed by a slow influx
5.3 Subpopulation Selection—When isolated from donor of water, as well as preservation agents, capable of penetrating
tissue, the populations of cells isolated may be heterogeneous. the cell membrane. The osmotic stresses resulting from intro-
Specific therapies or culture techniques may require the isola- duction (and removal) of a preservation solution can result in
tion of specific cell types (progenitor cells or mature cells of a lethal stresses to the cells. Protocols for introduction of
given phenotype). Methods of selecting these s
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